title: [Detection of foodborne Salmonella typhimurium and Salmonella enteritidis by diplex real-time PCR using TaqMan probe].
abstract: OBJECTIVE
  To establish a method to detect Salmonella typhimurium (ST) and Salmonella enteritidis (SE) simultaneously with a dual real-time PCR assay using double-color fluorescent TaqMan probes.
  
  
  METHODS
  The primers and probes were designed based on the conservative domain of STM4599 sequence of ST (GenBank: AERV01000023.1) and the specific sequence of SE ( GenBank: AF370707.1) respectively. The probes were labeled with reporter dye FAM for ST or VIC for SE at the 5' end. The dual real-time fluorescence PCR assay was set up and conditions were modified.
  
  
  RESULTS
  The dual real-time fluorescence PCR method for ST and SE was developed successfully. ST and SE specific primers and probes amplified 16 SE and 15 ST strains, while other 28 different Sa serotypes and 17 negative control Proteus strains showed negative results. The amplification efficiency of ST and SE with the dual fluorescent PCR were all 94. 2% and R2 were 0. 998 and 0. 995 respectively, while the minimum detectable concentration reached 300 CFU/ml for ST and 260 CFU/ml for SE. The entire test can be completed within 31 hours.
  
  
  CONCLUSIONS
  The method is highly specific, sensitive, and fast. The present study thus provides a rapid and effective method to detect ST and SE simultaneously from food samples.
authors.affiliation: Guangdong Inspection and Quarantine Technology Center, Guangzhou 510623, China. yuangmy@126.com
authors.forename: Muyun
authors.initials: M
authors.lastname: Yuan
authors.forename: Longyan
authors.initials: L
authors.lastname: Xu
authors.forename: Jijuan
authors.initials: J
authors.lastname: Cao
authors.forename: Jing
authors.initials: J
authors.lastname: Yang
authors.forename: Wang
authors.initials: W
authors.lastname: Zhang
authors.forename: Biling
authors.initials: B
authors.lastname: Chen
authors.forename: Dapeng
authors.initials: D
authors.lastname: Xiang
mesh_terms: D015342:DNA Probes; D004269:DNA, Bacterial; D005516:Food Microbiology; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D012477:Salmonella enteritidis / Q000302:isolation & purification*; D012486:Salmonella typhimurium / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D019914:Taq Polymerase / Q000737:chemistry / Q000235:genetics
chemical_list: D015342:DNA Probes; D004269:DNA, Bacterial; D019914:Taq Polymerase